Description of data: Statistical analysis of the nanozeolite internalization data for the different types of nanozeolites in the same time point and for the same type nanozeolite in different time points.
Interestingly, the analysis of the fMLP-induced internalization of the receptors in cells depleted of any of the components of the GIT complexes analysed showed a clear inhibition of the perinuclear accumulation of the internalized FPR.
The fluorescence images that we present, along with the biochemical biotinylation data, strongly support a massive internalization defect, consistent with the Boucrot et al. paper, in which the analysis of transferrin internalization was carried out by FACS in epsin triple knockdown cells.
For this reason, we did not deem it necessary to repeat the analysis of transferrin internalization using a biochemical assay.
To analyze sorting signals for the endocytic pathway, we have created GFP-based reporters that are post-translationally inserted into the PM but also allow the analysis of internalization signals by fusing the 26 C-terminal residues of AtROP10 (hereafter named Box) to the N-terminus of GFP.
Analysis of spore internalization in cells transfected with the different siRNAs indicated that knockdown of c-Src decreased spore internalization by approximately 60% (***, p<0.001 compared to the control), whereas knockdown of c-Yes or Fyn had no or only marginal inhibitory effect (Fig. 3, C).
Analysis of spore internalization showed that expression of Δp85α significantly decreased the spore internalization frequency (by ∼45%) in both A549 (Fig. 2, E, **, p<0.01) and HeLa cells (Fig. S1, D, ***, p<0.001) when compared to cells transfected with the vector control.
Another application of qPhage was the precise quantification of phage uptake in mammalian cells, thereby allowing analysis of its internalization dynamics.
As internalized molecules cannot be removed by cell washing or filtration, these cells contribute to the background level of fluorescence; however, they do not compromise our analysis of protein internalization.
A time course analysis of ethanol internalization showed that ethanol tissue concentration in 96 hpf larvae exposed to 350 mM ethanol approached 60 mM within 1 minute, peaked at 205 mM at 30 minutes and equilibrated to 140 mM by 1 hour, after which concentrations fluctuated between 150 and 175 mM (Fig. 3B).
